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Description
Mouse DAPK3 ELISA KitProduct Specification Usage Required experimental equipment: 1. Microplate reader (450nm) 2. High precision pipettes and pipette tips: 0. 5 10uL, 5 50uL, 20 200uL, 200 1000uL 3. 37C incubator 4. Distilled or deionized water Sample preparation and requirements: Tissue homogenization: Rinse the tissue with pre chilled PBS (0. 01M, pH 7. 4) to remove residual blood (lysed red blood cells in the homogenate will affect the measurement results). Weigh and
Product Specification
| Usage | Required experimental equipment:
1. Microplate reader (450nm) 2. High-precision pipettes and pipette tips: 0.5-10uL, 5-50uL, 20-200uL, 200-1000uL 3. 37°C incubator 4. Distilled or deionized water Sample preparation and requirements: Tissue homogenization: Rinse the tissue with pre-chilled PBS (0.01M, pH 7.4) to remove residual blood (lysed red blood cells in the homogenate will affect the measurement results). Weigh and mince the tissue. Add the minced tissue to the appropriate volume of PBS (generally a 1:9 weight-to-volume ratio, e.g., 1g of tissue sample to 9mL of PBS. The specific volume can be adjusted according to experimental needs and recorded. It is recommended to add protease inhibitors to the PBS) in a glass homogenizer and grind thoroughly on ice. To further lyse tissue cells, the homogenate can be ultrasonically disrupted or repeatedly frozen and thawed. Finally, the homogenate is centrifuged at 5000×g for 5-10 minutes and the supernatant is collected for analysis. Other biological fluids: Centrifuge at 1000xg for 20 minutes, remove the supernatant, and test. Pre-test preparation: 1. Remove the test kit from the refrigerator 10 minutes in advance and equilibrate to room temperature. 2. Prepare the standard gradient working solution: Add 1 mL of universal diluent to the lyophilized standard, let it stand for 15 minutes to completely dissolve, then gently mix (concentration is 10 ng/mL). Then dilute to the following concentrations: 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.3125 ng/mL, 0.15625 ng/mL, and 0 ng/mL. Serial dilution method: Take seven EP tubes and add 500uL of universal diluent to each. Pipette 500uL of the 10ng/mL standard working solution into the first EP tube and mix thoroughly to make a 5ng/mL standard working solution. Repeat this procedure for subsequent tubes. The last tube serves as a blank well; there is no need to pipette liquid from the penultimate tube. See the figure below for details. 3. Preparation of biotinylated detection antibody working solution: Centrifuge the concentrated biotinylated antibody at 1000×g for 1 minute 15 minutes before use. Dilute the 100× concentrated biotinylated antibody to a 1× working concentration with universal diluent (e.g., 10uL concentrate + 990uL universal diluent). Prepare and use immediately. 4. Prepare the enzyme conjugate working solution: 15 minutes before use, centrifuge the 100× concentrated enzyme conjugate at 1000×g for 1 minute. Dilute the 100× concentrated HRP enzyme conjugate to a 1× working concentration with universal diluent (e.g., 10 μL of concentrate + 990 μL of universal diluent). Prepare immediately. 5. Prepare the 1× wash solution: Dispense 10 mL of 20× wash solution into 190 mL of distilled water (concentrated wash solution removed from the refrigerator may crystallize; this is normal. Allow to stand at room temperature until the crystals have completely dissolved before preparing). Procedure: 1. Remove the desired strips from the aluminum foil bag after equilibration at room temperature for 10 minutes. Seal the remaining strips in a ziplock bag and return to 4°C. 2. Sample addition: Add 100 μL of sample or standard of varying concentrations to the corresponding wells. Add 100 μL of universal diluent to the blank wells. Cover with a film and incubate at 37°C for 60 minutes. (Recommendation: Dilute the sample to be tested at least 1-fold with universal diluent before adding it to the ELISA plate. This will reduce the impact of matrix effects on the test results. The sample concentration should be multiplied by the corresponding dilution factor when calculating the final sample concentration. It is recommended to run replicates for all test samples and standards.) 3. Add Biotinylated Antibody: Remove the ELISA plate and discard the liquid without washing. Add 100 μL of Biotinylated Antibody Working Solution directly to each well. Cover with a film and incubate at 37°C for 60 minutes. 4. Wash: Discard the liquid and add 300 μL of 1x Wash Solution to each well. Let stand for 1 minute, shake off the wash solution, and pat dry on absorbent paper. Repeat this process three times (a plate washer can also be used). 5. Add Enzyme Conjugate Working Solution: Add 100 μL of Enzyme Conjugate Working Solution to each well. Cover with a film and incubate at 37°C for 30 minutes. 6. Washing: Discard the liquid and wash the plate five times as in step 4. 7. Adding substrate: Add 90 μL of substrate (TMB) to each well, cover with a sealing film, and incubate at 37°C in the dark for 15 minutes. 8. Adding stop solution: Remove the ELISA plate and add 50 μL of stop solution directly to each well. Immediately measure the OD value of each well at a wavelength of 450 nm. Calculating experimental results: 1. Calculate the average OD value of the standard and sample replicates and subtract the OD value of the blank well as a correction factor. Plot the standard curve of the four-parameter logistic function on double-logarithmic graph paper, with concentration as the horizontal axis and OD value as the vertical axis. 2. If the sample OD value is higher than the upper limit of the standard curve, dilute the sample appropriately and retest. Multiply the sample concentration by the corresponding dilution factor. |
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| Theory | This kit utilizes a double-antibody sandwich enzyme-linked immunosorbent assay (ELISA). Sample, standard, biotin-labeled detection antibody, and HRP conjugate are sequentially added to microwells pre-coated with a Death Associated Protein Kinase 3 (DAPK3) capture antibody. After incubation and washing, the sample is developed using the substrate TMB. TMB is converted to blue by HRP peroxidase and to yellow by acid. The intensity of the color is positively correlated with the level of Death Associated Protein Kinase 3 (DAPK3) in the sample. The absorbance (OD) is measured at 450 nm using a microplate reader to calculate the sample concentration. | |||||||||||||||||||||||||||||||||
| Source | Mouse | |||||||||||||||||||||||||||||||||
| Synonym | Mouse Death Associated Protein Kinase 3 ELISA Kit | |||||||||||||||||||||||||||||||||
| Detection Type | Double antibody sandwich method | |||||||||||||||||||||||||||||||||
| Composition |
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| Background | Death-associated protein kinase 3, also known as DAPK3 or ZIPK, is an enzyme encoded by the DAPK3 gene. It is a member of the DAPK family. Death-associated protein kinases (DAPKs) are members of the serine/threonine protein kinase family that regulate cell death. When overexpressed in cells, they induce morphological changes consistent with apoptosis. They are involved in the regulation of apoptosis, autophagy, transcription, translation, and actin cytoskeletal reorganization. They are also involved in the regulation of smooth muscle contraction. They promote cancer cell proliferation rather than inducing apoptosis in various cancer cell lines. This gene regulates cell proliferation, migration, and invasion in A549 cells through ERK/c-Myc signaling. It has been shown to interact with PAWR and death-associated protein 6. | |||||||||||||||||||||||||||||||||
| General Notes | 1. Strictly adhere to the specified incubation time and temperature to ensure accurate results. All reagents must be at room temperature (20-25°C) before use. Refrigerate reagents immediately after use. 2. Improper plate washing may result in inaccurate results. Ensure that all liquid in the wells is aspirated thoroughly before adding substrate. Do not allow the wells to dry out during incubation. 3. Remove any residual liquid and fingerprints from the bottom of the plate, as this will affect the OD value. 4. The substrate developer solution should be colorless or very light in color. Do not use substrate solution that has turned blue. 5. Avoid cross-contamination of reagents and specimens to prevent erroneous results. 6. Avoid direct exposure to strong light during storage and incubation. 7. Do not expose any reagents to bleaching solvents or the strong fumes emitted by bleaching solvents. Any bleaching agent will destroy the biological activity of the reagents in the kit. 8. Do not use expired products, and do not mix components with different product numbers and batches. 9. Recombinant proteins from sources other than the kit may not be compatible with the antibodies in this kit and will not be recognized. 10. If there is a possibility of disease transmission, all samples should be managed properly and samples and testing devices should be handled according to prescribed procedures. |
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| Storage Temp. | If the unopened kit is stored at 4°C, the shelf life is 6 months. | |||||||||||||||||||||||||||||||||
| Test Range | 0.156-10 ng/mL | |||||||||||||||||||||||||||||||||
| Applications | Tissue homogenates and other biological fluids |
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4.2 ★★★★★
Based on 783 reviews
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The DXXULL 4-in-1 Aluminum Alloy Cigar Tube has quickly become my go-to accessory for enjoying cigars during my golf rounds. This sleek black tube combines multiple functions into one compact design, making it incredibly convenient for travel and daily use.
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-Cigar Holder: The tube comfortably holds a single cigar, protecting it from damage while on the move.
-Portable Design: Its compact size makes it easy to carry in my golf bag or pocket, ensuring I always have a cigar ready to enjoy.
The aluminum alloy construction feels sturdy and premium, and the airtight seal keeps my cigar in optimal condition. Whether I'm heading to the golf course or just out for a walk, this tube ensures my cigar is always ready for a relaxing smoke
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Reviewed in the United States on June 17, 2025
★★★★★ 5
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Reviewed in the United States on March 10, 2026
★★★★★ 5
What an incredible value and amazing quality!
When this came up, I had to get it! I have several differrent cigar cutters, punches, and other accessories, but none compare to this one! It feels like a high quality knife, but is a cigar tool that does it all. I thought it was weighted, but was only the quality steel and components I was feeling in my hand. I took it to the cigar store I go to and asked if they had seen anything like this. They said yes, but are usually $75+ when you find them. It opens in your palm with a simple squeeze between the fingers and palm. The steel feels like quality in your hand, and when you open it and see both a V and standard cutter as the main parts, it seems like that's all it does. Nope.....it also has a poker on one side and a circle punch that folds out on one end. It is so easy to use and is something you will want to show your cigar smoking friends, as well as sharing it with them. It works extremely well and is an incredible value at this price. Highly recommended for any cigar smoker!
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Reviewed in the United States on December 4, 2025
★★★★★ 5
Great looking cigar cutter, would make a nice gift
Nicely made and feels pretty good quality for the price. It's not heirloom quality or anything but it's very solid and made to last. Finish is great, it feels good in hand and has no sharp edges or blemishes as is common on many of these products.
This includes a pretty complete set of all the cutters you could need all in one device, would make either a nice starter tool or would be a great gift to upgrade to if you know anyone still using one of those cheap plastic ones from the cigar shop.
The steel appears to be chinese made and not super high quality, but it's also unlikely to see hard use. I would expect it to last most of a lifetime without sharpening if only used on clean cigars and not abused.
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Reviewed in the United States on December 17, 2025
★★★★★ 4
The Ultimate Cigar Toolkit: A 5-in-1 Must-Have for Aficionados
This 5-in-1 Cigar Cutter Set is an incredibly thoughtful and practical piece of gear, making it a fantastic gift for any cigar enthusiast—or a brilliant upgrade for yourself. While it's not a 'perfect 5-star' tool in every single function, its sheer versatility and excellent ergonomic design earn it a very strong four stars.
The main attraction is the built-in versatility. Having a Cigar V Cutter, Flat Cutter, and a Cigar Punch all in one compact, ergonomically-designed unit is a game-changer. It eliminates the need to carry three separate tools and the frustration of realizing you left your preferred cutter at home. The cutters themselves are sharp, providing clean, crisp cuts right out of the box, though, like any blade, their longevity will depend on regular maintenance.
Where this set truly shines and elevates itself beyond a simple cutter is the inclusion of the Cigar Draw Enhancer and the simple yet effective Cigar Holder. The draw enhancer is a lifesaver for those tightly packed cigars that refuse to cooperate; it's easy to use and significantly improves the smoking experience when needed. The holder is a small but welcome addition, giving you a clean place to rest your cigar when a proper ashtray isn't available.
The ergonomic design is well-executed. It feels substantial and secure in the hand, which is crucial for making a confident cut. It has a premium look and weight that makes it an impressive item to unbox and use.
The only reason this set doesn't quite hit a full five stars is minor: while all tools are functional, the punch and draw enhancer, while very useful, can occasionally be a little tricky to deploy or stow quickly compared to their standalone counterparts. For example, a dedicated V-cutter might offer a slightly deeper, more customized cut. However, this is an acceptable trade-off for the unparalleled convenience of having all five tools together.
Overall: If you value convenience, functionality, and portability, this 5-in-1 set is an excellent investment. It’s an ideal gift that says, "I want you to have the best experience possible with your cigars," and is genuinely useful on any occasion—whether it’s a Christmas stocking stuffer or a Father's Day present. You won't regret adding this versatile piece to your collection.
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Reviewed in the United States on October 15, 2025